Removal of the tag from His-tagged ILYd4, a human CD59 inhibitor, significantly improves its physical properties and its activity.
Removal of the tag from His-tagged ILYd4, a human CD59 inhibitor, significantly improves its physical properties and its activity.
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DOI：
10.2174/138161212802430486
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发表时间：
2012
影响因子：
3.1
通讯作者：
Qin X
中科院分区：
文献类型：
--
作者：
Wu L;Su S;Liu F;Xu T;Wang X;Huang Y;Sun X;Ge X;Chen T;Liu H;Wang C;Chorev M;Xu T;Qin X
补体依赖性细胞毒性（CDC）在治疗b细胞非霍奇金淋巴瘤（NHL）和慢性淋巴细胞白血病（CLL）中显著促进利妥昔单抗（RTX）和Ofatumumab （OFA）的疗效。人CD59 （hCD59）是一种关键的补体调节蛋白，它限制膜攻击复合物的形成，从而抑制CDC。hCD59是NHL和CLL对RTX和OFA治疗敏感性的重要决定因素。最近，我们开发了一种特异性和有效的hCD59抑制剂，his标记的ILYd4，它由30个氨基酸序列组成，从ILYd4的n端延伸。我们之前发表的结果表明，his标记的ILYd4可以作为进一步开发RTX和OFA治疗RTX耐药NHL和CLL的潜在治疗佐剂的主要候选物。然而，这些研究是使用在n端标记含有6 X His的30个额外氨基酸（AA）的ILYd4进行的，用于固定化金属亲和色谱仪。为了进一步开发基于ilyd4的治疗方法，我们研究了去除这一外来序列对抗hcd59活性的影响。在本文中，我们报道了无标签ILYd4的生成和表征。我们证明了无标签的ILYd4的抗hcd59活性比有his标签的ILYd4高3倍以上。rtx介导的CDC对b细胞恶性细胞的增强作用来自无标签ILYd4改善的功能和物理性质，包括更好的溶解度，降低聚集倾向和更高的热稳定性。因此，无标签ILYd4是进一步开发用于临床应用的更好候选物。
Complement dependent cytotoxicity (CDC) significantly contributes to Rituximab (RTX) and Ofatumumab (OFA) efficacies in the treatment of B-cell non-Hodgkin’s lymphoma (NHL) and chronic lymphocytic leukemia (CLL). Human CD59 (hCD59) is a key complement regulatory protein that restricts the formation of the membrane attack complex and thereby inhibits CDC. hCD59 is an important determinant of the sensitivity of NHL and CLL to RTX and OFA treatment. Recently, we developed a specific and potent hCD59 inhibitor, His-tagged ILYd4, which consists of 30 amino acid sequences extending from the N-terminus of ILYd4. Our previously published results indicate that His-tagged ILYd4 can be used as a lead candidate to further develop a potential therapeutic adjuvant for RTX and OFA treatment of RTX-resistant NHL and CLL. However, these studies were conducted using ILYd4 tagged on the N-terminus with 30 additional amino acids (AA) containing 6 X His used for immobilized metal affinity chromatograph. As a further step towards the development of ILYd4-based therapeutics, we investigated the impact of the removal of this extraneous sequence on the anti-hCD59 activity. In this paper, we report the generation and characterization of tag-free ILYd4. We demonstrate that tag-free ILYd4 has over three-fold higher anti-hCD59 activities than the His-tagged ILYd4. The enhanced RTX-mediated CDC effect on B-cell malignant cells comes from tag-free ILYd4’s improved functionality and physical properties including better solubility, reduced tendency to aggregation, and greater thermal stability. Therefore, tag-free ILYd4 is a better candidate for the further development for the clinical application.
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DOI：
10.1016/j.jchromb.2006.07.031
发表时间：
2006-12-05
影响因子：
3
作者：
Amor-Mahjoub, Mouna;Suppini, Jean-Philippe;Ladjimi, Moncef
通讯作者：
Ladjimi, Moncef
影响因子：
3.6
作者：
Gancz, Dana;Fishelson, Zvi
通讯作者：
Fishelson, Zvi
影响因子：
11.5
作者：
Ge, Xiaowen;Wu, Lin;Qin, Xuebin
通讯作者：
Qin, Xuebin
影响因子：
45.3
作者：
Bannerji, R;Kitada, S;Byrd, JC
通讯作者：
Byrd, JC
影响因子：
82.9
作者：
Hu, Weiguo;Ferris, Sean P.;Qin, Xuebin
通讯作者：
Qin, Xuebin